[摘 要] 目的：探讨DNA甲基转移酶1（DNMT1）通过稳定zeste基因增强子同源物2（EZH2）促进结直肠癌（CRC）HCT8细胞增殖与迁移的机制。方法：利用生物信息学方法分析DNMT1在CRC组织中的表达水平。WB法检测DNMT1在CRC细胞HCT8、SW620和正常结肠上皮细胞NCM460中的表达。通过siRNA或慢病毒载体转染HCT8细胞，分为siNC组、siDNMT1组、Vector组、DNMT1-OE组、siTRAF6组、siEZH2组、siEZH2 + DNMT1-OE组。采用克隆形成实验、CCK-8法、Transwell实验和划痕愈合实验检测敲低或过表达DNMT1对HCT8细胞增殖与迁移的影响，WB和qPCR法检测EZH2蛋白和mRNA水平，免疫沉淀（IP）法检测EZH2泛素化水平，免疫荧光双染检测肿瘤坏死因子受体相关因子6（TRAF6）与EZH2的细胞内共定位情况，克隆形成和划痕愈合实验验证EZH2对DNMT1功能的逆转作用。收集2022—2025年间郑州大学附属洛阳中心医院手术切除的12例CRC患者的癌及癌旁组织标本，采用免疫组化法检测CRC组织中DNMT1、TRAF6和EZH2的表达水平。结果：DNMT1在CRC组织中表达显著高于癌旁组织（P < 0.01），且在CRC细胞中表达上调（P < 0.05）；DNMT1敲低显著抑制HCT8细胞增殖及迁移（均P < 0.01），过表达则相反（均P < 0.01）。DNMT1正向调控EZH2的蛋白水平（P < 0.01），而mRNA水平不变（P > 0.05）。MG132可恢复siDNMT1组的EZH2蛋白表达（P < 0.01），且siDNMT1组EZH2泛素化水平升高。DNMT1负向调控TRAF6的表达（P < 0.01），且TRAF6与EZH2在细胞质中共定位，IP证实两者直接结合。敲低TRAF6可减弱EZH2的泛素化水平，敲低EZH2可逆转DNMT1对HCT8细胞增殖、迁移的促进作用（均P < 0.01）。DNMT1和EZH2在CRC组织中呈高表达（P < 0.01），TRAF6在CRC组织中表达显著低于癌旁组织（P < 0.05）。结论：DNMT1通过抑制TRAF6稳定EZH2促进CRC细胞的增殖和迁移，DNMT1、TRAF6和EZH2可能是CRC治疗的潜在靶点。

ObjectiveTo establish a model of dry eye with lung Yin deficiency syndrome in mice. MethodsA total of 40 SPF C57BL/6J mice were assigned via the random number table method into 5 groups （n=8）： Normal control， model control， and high-， medium-， and low-dose （11.7， 5.85， and 2.925 g·kg^-1， respectively） Yangyin Qingfeitang. Mice in the normal control group were fed normally without any intervention. Mice in Yangyin Qingfeitang group and model control group were treated with 0.2% benzalkonium chloride eye drops （5 μL） twice a day and fed in a controlled drying system in a dry environment for 28 days. At the same time， the mice were administrated with thyroxine tablet solution by gavage and placed in a glass fumigation tank （SO₂ concentration： 0.5 g·m^-3） for 14 days. After 4 weeks， mice in Yangyin Qingfeitang groups were treated with Yangyin Qingfeitang by gavage and those in the normal control group and model control group were administrated with deionized water at 0.01 mL·g^-1. The body mass， anal temperature， four examination information （claw and nail appearance）， basic tear secretion test， tear film rupture time， corneal fluorescein staining， and lacrimal gland HE staining were compared among groups. Compound Yangyin Qingfeitang granules were used to measure the syndrome to verify the success of modeling. ResultsAfter 28 days of continuous modeling， compared with the normal control group， the model group exhibited listless and emaciated status， coughing， drowsiness， dry and dull hair， dry and hard stool， reduced food intake and water intake， red lip circumference， red tongue with reduced fluid， dry nose and teeth， red claws and nails， body mass gain， decreased anal mild tear secretion （P<0.05）， and shortened tear film rupture time （P<0.05）. After 28 days of modeling， the mice showed large corneal fluorescein staining range， severe corneal injury， and increased content of interleukin （IL）-18， IL-β， and tumor necrosis factor （TNF）-α in lacrimal gland， compared with those in the normal control group （P<0.05）. After the treatment with Yangyin Qingfeitang， the mice had good drinking and eating conditions， with lighter redness of the tongue， moist nose， moist and shiny teeth， and the claw and nail color close to that in the normal group. Compared with the model control group， Yangyin Qingfeitang groups showed increases in body mass and anal temperature （P<0.05）， tear secretion （P<0.05）， and tear film rupture time （P<0.05）， narrowed range of corneal fluorescein staining， and declined levels of IL-18， IL-β， and TNF-α in lacrimal glands （P<0.01）. The high-dose group had the best effect， with the indicators close to the levels in the normal control group. ConclusionThe animal model of dry eye with lung Yin deficiency syndrome can be established by culture in a controlled drying system， treatment with benzalkonium chloride eye drops for 28 days， and administration of thyroxine tablet solution combined with SO₂ fumigation for 14 days.

ObjectiveTo investigate the mechanisms of Runmu Dihuang decoction （RMDHD） in treating perimenopausal dry eye with liver-kidney Yin deficiency syndrome based on the silent information regulator 3 （SIRT3）/hypoxia-inducible factor-1α （HIF-1α）/nuclear factor-κB （NF-κB） signaling pathway. MethodsSixty female Sprague-Dawley rats were randomly divided into six groups （n=10 per group）： Sham operation group， model group， sodium hyaluronate eye drop group， and low-， medium-， and high-dose RMDHD groups （5.625， 11.25， 22.50 g·kg^-1）. Except for the sham operation group， all rats underwent bilateral ovariectomy and were administered 0.1% benzalkonium chloride eye drops combined with long-term chronic irritation to establish a perimenopausal dry eye model with liver-kidney Yin deficiency syndrome. Drug administration began in the 11th week after modeling and continued for 21 days. General conditions， screen-grip test scores， tear secretion volume， tear film breakup time （TFBUT）， and corneal fluorescein staining were recorded. Serum levels of reactive oxygen species （ROS）， follicle-stimulating hormone （FSH）， estradiol （E₂）， and progesterone （PROG） were measured by enzyme-linked immunosorbent assay （ELISA）. Pathological changes in the lacrimal glands， corneas， and uteri were observed using hematoxylin-eosin （HE） staining. Protein expression levels of SIRT3， HIF-1α， phosphorylated NF-κB p65 （p-NF-κB p65）， and total NF-κB p65 in the lacrimal glands were detected by Western blot. The expression of inflammatory cytokines interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） in the lacrimal glands was assessed by immunohistochemistry （IHC）. ResultsAfter model establishment， no significant differences were observed among the groups except the sham operation group. Compared with the sham operation group， the other groups exhibited slowed movement， dull responses， increased irritability， reduced body weight， elevated rectal temperature， decreased screen-grip test scores， reduced tear secretion， and significantly shortened TFBUT （P<0.05）. After treatment， compared with the model group， the sodium hyaluronate eye drop group and all RMDHD groups showed improved general conditions， significantly increased tear secretion （P<0.05）， prolonged TFBUT （P<0.05）， and elevated screen-grip test scores （P<0.05）. Serum ROS and FSH levels were significantly decreased， while E₂ and PROG levels were significantly increased （P<0.05）. Pathological damage to the cornea， lacrimal glands， and uterus was ameliorated. In addition， protein expression levels of SIRT3 and HIF-1α in the lacrimal glands were significantly upregulated （P<0.05）， whereas the expression of p-NF-κB p65， IL-1β， and TNF-α was significantly downregulated （P<0.05）. ConclusionRMDHD increases tear secretion and TFBUT， improves lacrimal gland and corneal injury， and alleviates dry eye symptoms in a perimenopausal dry eye rat model with liver-kidney Yin deficiency syndrome. The underlying mechanism may be related to regulation of the SIRT3/HIF-1α/NF-κB signaling pathway， inhibition of oxidative stress and inflammatory responses， and reduction of ocular surface tissue damage.

BackgroundChronic insomnia disorder has become a significant public health issue， and it may be associated with dyslipidemia. Previous studies on dyslipidemia in patients with chronic insomnia disorder have mainly focused on exploring the relationship between subjective short sleep duration and dyslipidemia， while there have been limited studies on the relationship between objective short sleep duration and dyslipidemia. ObjectiveTo explore the relationship between objective short sleep duration and dyslipidemia in patients with chronic insomnia disorder， in order to provide references for the prevention and intervention of dyslipidemia in this population. MethodsA total of 103 patients who were hospitalized at The Third Hospital of Mianyang from August 2022 to November 2023 and met the diagnostic criteria for chronic insomnia disorder as defined in the International Classification of Sleep Disorder， third edition （ICSD-3） were retrospectively collected. The objective sleep duration of the patients was obtained through polysomnography. The patients were divided into two groups based on their objective sleep duration： the group with objective sleep duration ≥ 7 hours （n=71） and the group with objective sleep duration < 7 hours （n=32）. Binary Logistic regression analysis was used to explore the impact of objective sleep duration < 7 hours on dyslipidemia. ResultsAmong 103 patients with chronic insomnia disorder， 59 cases （57.28%） were identified with dyslipidemia. The comparison of dyslipidemia conditions between the group with objective sleep duration ≥ 7 hours and the group with objective sleep duration < 7 hours showed a statistically significant difference （χ²=5.956， P<0.05）. Compared with the group with objective sleep duration ≥7 hours， the group with objective sleep duration < 7 hours exhibited significantly lower high-density lipoprotein cholesterol levels， and reduced sleep efficiency （t=-2.003， -5.482， P<0.05 or 0.01）. Binary Logistic regression analysis results showed that the risk of abnormal blood lipids in patients with chronic insomnia disorder with objective sleep duration < 7 hours was 3.128 times higher than that of patients with objective sleep duration ≥ 7 hours （OR=3.128， 95% CI： 1.139–8.588）. ConclusionObjective short sleep duration may be a risk factor for dyslipidemia in patients with chronic insomnia disorder.

ObjectiveUlcerative colitis is a prevalent immunoinflammatory disease. Th17/Treg cell imbalance and gut microbiota dysregulation are key factors in ulcerative colitis pathogenesis. The actin cytoskeleton contributes to regulating the proliferation, differentiation, and migration of Th17 and Treg cells. Wdr63, a gene containing the WD repeat domain, participates in the structure and functional modulation of actin cytoskeleton. Recent research indicates that WDR63 may serve as a regulator of cell migration and metastasis via actin polymerization inhibition. This article aims to explore the effect of Wdr63 deletion on Th17/Treg cells and ulcerative colitis. MethodsWe constructed Wdr63^-/- mice, induced colitis in mice using dextran sulfate sodium salt, collected colon tissue for histopathological staining, collected mesenteric lymph nodes for flow cytometry analysis, and collected healthy mouse feces for microbial diversity detection. ResultsCompared with wild-type colitis mice, Wdr63^-/- colitis mice had a more pronounced shortening of colonic tissue, higher scores on disease activity index and histological damage index, Treg cells decreased and Th17 cells increased in colonic tissue and mesenteric lymph nodes, a lower level of anti-inflammatory cytokine IL-10, and a higher level of pro-inflammatory cytokine IL-17A. In addition, WDR63 has shown positive effects on maintaining intestinal microbiota homeostasis. It maintains the balance of Bacteroidota and Firmicutes, promoting the formation of beneficial intestinal bacteria linked to immune inflammation. ConclusionWdr63 deletion aggravates ulcerative colitis in mice, WDR63 inhibits colonic inflammation likely by regulating Th17/Treg balance and maintains intestinal microbiota homeostasis.

Objective To investigate the role and mechanism of ubiquitin-specific protease 46 (USP46) in hypoxia/reoxygenation (H/R)-induced pyroptosis of renal tubular epithelial cells. Methods Renal tubular epithelial cells were divided into negative control siRNA group (si-CTL group), USP46 knockdown group (si-USP46 group), negative control siRNA + H/R treatment group (si-CTL+H/R group), and USP46 knockdown + H/R treatment group (si-USP46+H/R group). Flow cytometry was used to detect cell apoptosis in each group. Real-time quantitative polymerase chain reaction (RT-qPCR) was employed to measure the messenger RNA (mRNA) expression of USP46, NOD-like receptor protein 3 (NLRP3), gasdermin D (GSDMD), interleukin (IL)-18, and IL-1β. Western blotting was used to detect the protein expression of USP46, NLRP3, GSDMD, and cleaved cysteinyl aspartate specific proteinase (C-Caspase)-1. The levels of inflammatory factors and lactate dehydrogenase (LDH) in the cell supernatants were detected, and the levels of reactive oxygen species (ROS) and malondialdehyde (MDA) in the cells were detected. Co-immunoprecipitation was used to verify the interaction between USP46 and NLRP3. Results Compared with the si-CTL group, the si-CTL+H/R group exhibited increased cell apoptosis, elevated protein expression of USP46, NLRP3, GSDMD-N and C-Caspase-1, increased mRNA expression of USP46, NLRP3, GSDMD, IL-18 and IL-1β, higher levels of IL-18, IL-1β, TNF-α and LDH, and increased ROS and MDA levels (all P < 0.05). Compared with the si-CTL+H/R group, the si-USP46+H/R group showed decreased cell apoptosis, reduced protein expression of USP46, NLRP3, GSDMD-N and C-Caspase-1, decreased mRNA expression of USP46, GSDMD and IL-18, lower levels of IL-18, IL-1β, TNF-α and LDH, and decreased ROS and MDA levels (all P < 0.05). Co-immunoprecipitation results indicated that USP46 could bind to NLRP3. Conclusions Downregulation of USP46 alleviates H/R-induced pyroptosis in renal tubular epithelial cells, possibly by inhibiting USP46-dependent NLRP3 deubiquitination and promoting NLRP3 ubiquitination and degradation.

Objective: The existence of activating transcription factor 1 (ATF1) could be employed as a clinical marker in the context of cervical cancer development, although its specific mechanism has not been fully clarified. Methods: To evaluate the presence of ATF1, miR-630, and myelin and lymphocyte protein 2 (MAL2) in cervical malignancies, we conducted quantitative reverse transcription polymerase chain reaction, immunohistochemistry, and Western blot assays; further studied the expansion, migration, invasion and epithelial-mesenchymal transition (EMT) of cervical carcinoma cells using colony formation assay, transwell, loss cytometry, Western blot.Chromatin immunoprecipitation (ChIP) and RNA immunoprecipitation (RIP) were used to verify that ATF1 could directly transcriptionally repress miR-630; dual luciferase reporter assay and RIP assay were employed to confirm that miR-630 targeted to repress MAL2. Results: In cervical cancer cases, elevated ATF1 expression and reduced miR-630 expression were detected, displaying a negative relationship between them. Inhibition of ATF1 hindered the growth, migration, infiltration, and EMT in cervical carcinoma cells, while upregulation of miR-630 mitigated the aggressive characteristics of these cells. ATF1 was found to transcriptionally repress miR-630 by TransmiR and ALGGEN prediction and ChIP validation.MicroRNA modulates gene expression and affects cancer progression, and we discovered that miR-630 regulates cancer progression by targeting and inhibiting MAL2. Conclusion ATF1, which modulates the miR-630/MAL2 pathway, affects the EMT process and cervical carcinoma cell growth and spread. Therefore, ATF1 may serve as a promising marker and treatment target for cervical malignancies intervention.

Objective: The existence of activating transcription factor 1 (ATF1) could be employed as a clinical marker in the context of cervical cancer development, although its specific mechanism has not been fully clarified. Methods: To evaluate the presence of ATF1, miR-630, and myelin and lymphocyte protein 2 (MAL2) in cervical malignancies, we conducted quantitative reverse transcription polymerase chain reaction, immunohistochemistry, and Western blot assays; further studied the expansion, migration, invasion and epithelial-mesenchymal transition (EMT) of cervical carcinoma cells using colony formation assay, transwell, loss cytometry, Western blot.Chromatin immunoprecipitation (ChIP) and RNA immunoprecipitation (RIP) were used to verify that ATF1 could directly transcriptionally repress miR-630; dual luciferase reporter assay and RIP assay were employed to confirm that miR-630 targeted to repress MAL2. Results: In cervical cancer cases, elevated ATF1 expression and reduced miR-630 expression were detected, displaying a negative relationship between them. Inhibition of ATF1 hindered the growth, migration, infiltration, and EMT in cervical carcinoma cells, while upregulation of miR-630 mitigated the aggressive characteristics of these cells. ATF1 was found to transcriptionally repress miR-630 by TransmiR and ALGGEN prediction and ChIP validation.MicroRNA modulates gene expression and affects cancer progression, and we discovered that miR-630 regulates cancer progression by targeting and inhibiting MAL2. Conclusion ATF1, which modulates the miR-630/MAL2 pathway, affects the EMT process and cervical carcinoma cell growth and spread. Therefore, ATF1 may serve as a promising marker and treatment target for cervical malignancies intervention.

Objective: The existence of activating transcription factor 1 (ATF1) could be employed as a clinical marker in the context of cervical cancer development, although its specific mechanism has not been fully clarified. Methods: To evaluate the presence of ATF1, miR-630, and myelin and lymphocyte protein 2 (MAL2) in cervical malignancies, we conducted quantitative reverse transcription polymerase chain reaction, immunohistochemistry, and Western blot assays; further studied the expansion, migration, invasion and epithelial-mesenchymal transition (EMT) of cervical carcinoma cells using colony formation assay, transwell, loss cytometry, Western blot.Chromatin immunoprecipitation (ChIP) and RNA immunoprecipitation (RIP) were used to verify that ATF1 could directly transcriptionally repress miR-630; dual luciferase reporter assay and RIP assay were employed to confirm that miR-630 targeted to repress MAL2. Results: In cervical cancer cases, elevated ATF1 expression and reduced miR-630 expression were detected, displaying a negative relationship between them. Inhibition of ATF1 hindered the growth, migration, infiltration, and EMT in cervical carcinoma cells, while upregulation of miR-630 mitigated the aggressive characteristics of these cells. ATF1 was found to transcriptionally repress miR-630 by TransmiR and ALGGEN prediction and ChIP validation.MicroRNA modulates gene expression and affects cancer progression, and we discovered that miR-630 regulates cancer progression by targeting and inhibiting MAL2. Conclusion ATF1, which modulates the miR-630/MAL2 pathway, affects the EMT process and cervical carcinoma cell growth and spread. Therefore, ATF1 may serve as a promising marker and treatment target for cervical malignancies intervention.

BackgroundPostpartum depression can affect the physical and mental health of mothers and the quality of parenting. Most Chinese women perform traditional postpartum practices （commonly known as "doing the month"） after giving birth， while the existing findings are inconsistent and inconclusive regarding the potential of traditional Chinese postpartum practices to alleviate or exacerbate postpartum depression. ObjectiveTo explore the relationship between traditional Chinese postpartum practices and postpartum depression， so as to provide references for reducing the risk of postpartum depression. MethodsA total of 240 consecutive women who gave birth in the obstetrics department of the Mianyang Central Hospital and the Third Hospital of Mianyang from January to May 2024 were selected. Data were collected using Self-designed General Information Questionnaire， Chinese version of the Edinburgh Postnatal Depression Scale （EPDS）， the Social Support Rating Scale （SSRS）， the Patient Health Questionnaire-15 （PHQ-15）， the Adherence to Doing-the-Month Practices questionnaire （ADP）， and the Self-compiled Questionnaire on the Cognition of Doing-the-Month. The absolute value （A value） of the difference between scores of ADP and Cognition of Doing-the-Month Questionnaire was calculated to evaluate the degree of cognitive behavioral conflict of postpartum women. Pearson correlation analysis was performed to examine the correlations of EPDS score with SSRS score， PHQ-15 score， ADP total and dimensional scores， Cognition of Doing-the-Month Questionnaire total and dimensional scores， and A value. Logistic regression analysis was conducted to identify the protective and risk factors for developing postpartum depression. ResultsThe postpartum depression was detected in 22.50% of women. The postpartum women had a EPDS score of （6.21±5.00）， ADP score of （70.05±20.57）， SSRS score of （41.96±6.96）， PHQ-15 score of （4.63±3.77）， and Cognition of Doing-the-Month questionnaire score of （40.30±10.13）. The A value was （0.65±0.58）. Correlation analysis revealed that EPDS score was negatively correlated with the total ADP score and the four dimensional scores of the restrictions on social activities， diet， housework， and personal hygiene （r=-0.228， -0.146， -0.184， -0.275， -0.168， P<0.05 or 0.01）， and positively correlated with the A value （r=0.161， P<0.05）. Logistic regression analysis indicated that restriction on housework dimension in ADP was entered into the model （OR=0.930， 95% CI： 0.885~0.978）. ConclusionThe restriction on housework dimension in traditional Chinese postpartum practices may be a protective factor against postpartum depression.